hsaec lung epithelial cells  (ATCC)

Journal: Frontiers in Microbiology

Article Title: Nitric oxide as a regulator of B. anthracis pathogenicity

doi: 10.3389/fmicb.2015.00921

Figure Lengend Snippet: Cytotoxic effect of Sterne and ΔNOS static culture sups. Bacteria were grown in DMEM (10 ml, 24 h, 37°C, 5% CO 2 ) in static conditions and the culture sups were prepared. The monolayers of human small-airway lung epithelial cells (HSAECs) were exposed to the bacterial sups for 2 h and the viability of the cells was determined using Resazurin dye as described in Section “Materials and Methods”. The final pH values of the sups and control medium are shown. Asterisks indicate the sups titrated to the pH 5.2 of Sterne sup after cultivation. The error bars indicate 95% confidence intervals of triplicate measurements.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) from Cambrex, Inc., Walkersville, MD, USA were used for viability and permeability assays.

Techniques: Bacteria, Control

Journal: Frontiers in Microbiology

Article Title: Nitric oxide as a regulator of B. anthracis pathogenicity

doi: 10.3389/fmicb.2015.00921

Figure Lengend Snippet: Cell-permeabilizing effects of Sterne and ΔNOS static culture sups. (A) Toxicity of the Sterne sup toward lung epithelial cells (HSAECs) correlates with increased permeability. HSAECs were exposed to the static culture sups for the indicated periods of time and the viability and permeability were tested as described in Section “Materials and Methods”. (B) Comparative effect of ΔNOS and Sterne sups after a 30-min exposure of HSAECs in conditions equal to (A) . Cholesterol added to the culture medium reveals presence of the pH-dependent permeabilizing factor(s). Asterisks indicate the sups and medium titrated to the pH 5.1 of Sterne sup after cultivation. Error bars show 95% confidence intervals of triplicate measurements.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) from Cambrex, Inc., Walkersville, MD, USA were used for viability and permeability assays.

Techniques: Permeability

Journal: Frontiers in Microbiology

Article Title: Nitric oxide as a regulator of B. anthracis pathogenicity

doi: 10.3389/fmicb.2015.00921

Figure Lengend Snippet: Effect of Sterne and ΔNOS sups grown in microaerobic conditions on phosphorylation of AKT (S473) in HSAECs. The cells were exposed for 2 h to the sups of bacterial cultures grown for 24 h in static conditions. Corresponding pH values of the sups are shown. The lysates of the cells were used for western blots with a primary rabbit antibody against phosphorylated form of AKT (S473) and a secondary anti-rabit antibody conjugated with horseradish peroxidase. The activity of peroxidase in the bands was detected using a SuperSignal West Femto chemiluminescent substrate (Pierce) and quantitated with a digital camera. To take into account the effect of pH the ΔNOS sup after cultivation was titrated from pH 6.4 to pH 5.1 indicated by an asterik. Error bars show 95% confidence intervals obtained from three independent western blots. The preparation of the lysates was repeated twice with similar results.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) from Cambrex, Inc., Walkersville, MD, USA were used for viability and permeability assays.

Techniques: Phospho-proteomics, Western Blot, Activity Assay